title: The establishment and validation of efficient assays for anti-IIa and anti-Xa activities of heparin sodium and heparin calcium.
abstract: Heparin is used as an anticoagulant drug. The anticoagulation process is mainly caused by the interaction of heparin with antithrombin followed by inhibition of anticoagulant factor IIa and factor Xa. The anti-factor IIa and anti-factor Xa activities of heparin are critical for its anticoagulant effect; however, physicochemical methods that can reflect these activities have not been established. Thus, the measurements of anti-IIa and anti-Xa activities by biological assay are critical for the quality control of heparin products. Currently in the Japanese Pharmacopoeia (JP), the activities of heparin sodium and heparin calcium are measured by an anti-Xa activity assay (anti-Xa assay), but anti-IIa activity is not measured. Here, we established an anti-IIa activity assay (anti-IIa assay) and an anti-Xa assay having good accuracy and precision. When samples having a relative activity of 0.8, 1.0 and 1.2 were measured by the established anti-IIa and anti-Xa assays in nine laboratories, good accuracy (100.0-102.8% and 101.6-102.8%, respectively), good intermediate precision (1.9-2.1% and 2.4-4.2%, respectively) and good reproducibility (4.0-4.8% and 3.6-6.4%, respectively) were obtained. The established anti-IIa and anti-Xa assays have similar protocols, and could be performed by a single person without a special machine. The established assays would be useful for quality control of heparin.
authors.affiliation: National Institute of Health Sciences, Japan. Electronic address: tsuzuki@nihs.go.jp.
authors.forename: Takuo
authors.initials: T
authors.lastname: Suzuki
authors.forename: Akiko
authors.initials: A
authors.lastname: Ishii-Watabe
authors.forename: Noritaka
authors.initials: N
authors.lastname: Hashii
authors.forename: Yukari
authors.initials: Y
authors.lastname: Nakagawa
authors.forename: Tomoko
authors.initials: T
authors.lastname: Takahashi
authors.forename: Akiko
authors.initials: A
authors.lastname: Ebisawa
authors.forename: Seiichi
authors.initials: S
authors.lastname: Nishi
authors.forename: Naho
authors.initials: N
authors.lastname: Fujita
authors.forename: Aya
authors.initials: A
authors.lastname: Bando
authors.forename: Yuko
authors.initials: Y
authors.lastname: Sekimoto
authors.forename: Kazuyoshi
authors.initials: K
authors.lastname: Miyata
authors.forename: Toshio
authors.initials: T
authors.lastname: Endo
authors.forename: Takuma
authors.initials: T
authors.lastname: Otsu
authors.forename: Shiori
authors.initials: S
authors.lastname: Sugimoto
authors.forename: Tadashi
authors.initials: T
authors.lastname: Kondou
authors.forename: Yuji
authors.initials: Y
authors.lastname: Fujita
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Miyanaga
authors.forename: Masahiro
authors.initials: M
authors.lastname: Mashimo
authors.forename: Norihiko
authors.initials: N
authors.lastname: Shimada
authors.forename: Hikaru
authors.initials: H
authors.lastname: Yoden
authors.forename: Hideo
authors.initials: H
authors.lastname: Shimamura
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Kurata
authors.forename: Sadatoshi
authors.initials: S
authors.lastname: Koyama
authors.forename: Nana
authors.initials: N
authors.lastname: Kawasaki
mesh_terms: D000925:Anticoagulants / Q000378:metabolism / Q000494:pharmacology; D000991:Antithrombins / Q000378:metabolism; D015951:Factor Xa / Q000378:metabolism; D065427:Factor Xa Inhibitors; D006493:Heparin / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D009842:Oligopeptides / Q000378:metabolism; D011516:Prothrombin / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015203:Reproducibility of Results; D013678:Technology, Pharmaceutical / Q000379:methods
chemical_list: D000925:Anticoagulants; D000991:Antithrombins; D065427:Factor Xa Inhibitors; D009842:Oligopeptides; C006871:calcium heparin; C015608:benzoyl-Ile-Glu-Gly-Arg-p-nitroanilide; D011516:Prothrombin; C025337:Factor IIa; D006493:Heparin; D015951:Factor Xa
keywords: Anti-IIa assay; Anti-Xa assay; Heparin; Quality control